Overview

  • Product name
    Anti-PDI antibody [RL90]
    See all PDI primary antibodies
  • Description
    Mouse monoclonal [RL90] to PDI
  • Tested applications
    Suitable for: ICC/IF, Electron Microscopy, IHC-P, IHC-Fr, IP, WB, ELISA, Inhibition Assay, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Hamster, Dog, Human, Pig, Monkey, African green monkey
    Predicted to work with: Drosophila melanogaster
  • Immunogen

    Other Immunogen Type corresponding to Rat PDI. Purified rat PDI protein.

  • Positive control
    • rat liver

Properties

Applications

Our Abpromise guarantee covers the use of ab2792 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/100. PubMed: 17308099
Electron Microscopy Use at an assay dependent concentration. PubMed: 21886772
IHC-P 1/100. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
IHC-Fr 1/100.
IP Use at an assay dependent concentration. This antibody has been shown to inhibit the activity of PDI in vitro. It has also been found to inhibit disulfide bond reduction of the HIV protein, gp120, at the cell surface of CHO cells and human lymphoid cells.
WB 1/1000. Detects a band of approximately 59-61 kDa (predicted molecular weight: 58 kDa). If there is no signal or signal is weak, more concentrated antibody could be used in addition to using less stringent blocking conditions (e.g., BSA instead of milk, incubating the antibody in PBST or TBST only, lower milk percentage).
ELISA Use at an assay dependent concentration.
Inhibition Assay Use at an assay dependent concentration.
Flow Cyt Use 0.5µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Acts as an intracellular estrogen-binding protein. May be involved in modulating cellular levels and biological functions of estrogens in the pancreas. May act as a chaperone that inhibits aggregation of misfolded proteins.
  • Tissue specificity
    Highly expressed in pancreas (at protein level).
  • Sequence similarities
    Belongs to the protein disulfide isomerase family.
    Contains 2 thioredoxin domains.
  • Post-translational
    modifications
    The disulfide-linked homodimer exhibits an enhanced chaperone activity.
    Glycosylated.
  • Cellular localization
    Endoplasmic reticulum lumen.
  • Information by UniProt
  • Database links
  • Alternative names
    • Pancreas specific protein disulfide isomerase antibody
    • Pancreas-specific protein disulfide isomerase antibody
    • Pancreatic protein disulfide isomerase antibody
    • PDA2 antibody
    • PDI antibody
    • PDIA2 antibody
    • PDIA2_HUMAN antibody
    • PDIp antibody
    • PDIR antibody
    • Protein disulfide isomerase A2 antibody
    • Protein disulfide isomerase antibody
    • Protein disulfide isomerase associated 2 antibody
    • Protein disulfide isomerase family A member 2 antibody
    • Protein disulfide isomerase pancreatic antibody
    • Protein disulfide-isomerase A2 antibody
    • Rho GDP dissociation inhibitor gamma antibody
    see all

Anti-PDI antibody [RL90] images



  • Predicted band size : 58 kDa
    Western blot analysis of PDI was performed by loading 25 ug of HepG2 (Lane 1) Hela (Lane 2) and NIH-3T3 (Lane 3) cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4?C overnight. The membrane was probed with ab2792 at 1:1000 overnight at 4?C and washed in TBST. The membrane was then probed with a HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using a ECL Plus Western Blotting Substrate. Results show a band at approx. 57 kDa.
  • Immunocytochemistry/Immunofluorescence analysis of PDI (red) in U2OS cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with ab2792 (1:75) for at least 1 hour at room temperature and incubated with DyLight 550 goat anti-mouse IgG secondary antibody (1:250) for 30 minutes at room temperature. Actin was stained with DyLight 488 Phalloidin at a dilution of 1:300 (1 unit/ml final concentration) for 30 minutes. Images were taken at 20X magnification.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human colon tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing PDI ab2792 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Overlay histogram showing HeLa cells stained with ab2792 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2792, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Anti-PDI antibody [RL90] (ab2792) at 1/2000 dilution

    Secondary
    Donkey anti mouse IgG2a at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 58 kDa
    Observed band size : 57 kDa (why is the actual band size different from the predicted?)


    Exposure time : 20 seconds

    This image is courtesy of an anonymous Abreview

    See Abreview



  • Predicted band size : 58 kDa

    See Abreview

  • ab2792 staining PDI in MDA MB 231 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 1% Triton X-100 and blocked with 10% BSA for 1 hour at 21°C. Samples were incubated with primary antibody (1/100 in BSA + 0.02% Tween 20) for 1 hour at 16°C. A DyLight® 550-conjugated goat anti-mouse IgG polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • Immunocytochemistry/Immunofluorescence analysis of PDI (green) in NIH 3T3 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with ab2792 (1:75) for at least 1 hour at room temperature and incubated with Dylight 488 goat anti-mouse IgG secondary antibody (1:250) for 30 minutes at room temperature. Actin was stained with Dylight 350 Phalloidin at a dilution of 1:120 (2.5units/ml final concentration) and nuclei (red) were stained with DRAQ5 at a concentration of 1ug/ml for 30 minutes. Images were taken at 20X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of PDI (green) in NIH 3T3 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were incubated with ab2792 (1:75) for at least 1 hour at room temperature and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (1:250) for 30 minutes at room temperature. Actin was stained with DyLight 550 Phalloidin at a dilution of 1:120 (2.5 units/ml final concentration) and nuclei (blue) were stained with Hoechst at a concentration of 1ug/ml for 30 minutes. Images were taken at 20X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of PDI (green) in NIH 3T3 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were incubated with ab2792 (1:75) for at least 1 hour at room temperature and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (1:250) for 30 minutes at room temperature. Actin was stained with DyLight 650 Phalloidin at a dilution of 1:120 (2.5 units/ml final concentration) and nuclei (blue) were stained with Hoechst at a concentration of 1ug/ml for 30 minutes. Images were taken at 20X magnification.

  • Immunocytochemistry/Immunofluorescent analysis of PDI using ab2792 shows staining in p19 Cells.
  • Immunocytochemistry/Immunofluorescent analysis of PDI using ab2792 shows staining in NS-1 Cells.
  • Immunocytochemistry/Immunofluorescent analysis of PDI using ab2792 shows staining in HMVEC Cells.
  • Immunocytochemistry/Immunofluorescent analysis of PDI using ab2792 shows staining in A549 Cells.
  • ab2792 positively staining dog MDCK cell ER (red) at 1/200. Staining was carried out in conjunction with goat anti mouse H and L ( Alexa 546) at 1/1000. The nuclei can be seen stained with Hoechst (blue)
    This image is courtesy of an Abreview submitted by Kun Liu on 20 September 2005. We do not have any further information relating to this image.

     

     

  • ab2792 positively staining formaldehyde fixed human Hek293 cells (1/200). This Ab was used in conjunction with goat anti mouse Alexa Fluor® 546 (1/1500). The nuclei has been atained with Hoechst.
    This image is courtesy of an Abreview submitted by Kun Liu on 19th September 2005. We do not have any further information relating to this image.

    See Abreview

  • ab2792 staining PDI from human HaCaT keratinocyte cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with paraformaldehyde, permeabilized with 0.25 % Triton ×100 and blocking with 2.5% BSA plus 1% goat serum for 1 hour at 40C was performed. Samples were incubated with primary antibody, diluted 1/100, for 1 hour at 240C. An Alexa Fluor ® 488-conjugated goat polyclonal to mouse IgG was used at dilution at 1/1000 as secondary antibody.

    See Abreview

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human tonsil tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing PDI ab2792 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human lung adenocarcinoma tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing PDI ab2792 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Flow cytometry analysis of PDI showing positive staining in the membrane and cytoplasm of K562 cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2792 at 1 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
  • Flow cytometry analysis of PDI showing positive staining in the membrane and cytoplasm of NIH/3T3 cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2792 at 0.5 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
  • Flow cytometry analysis of PDI showing positive staining in the membrane and cytoplasm of Hela cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2792 at 1 ug/test for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.

References for Anti-PDI antibody [RL90] (ab2792)

This product has been referenced in:

See all 95 Publications for this product

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I am glad to hear that the results with ab2792 were excellent.

I regret to inform you that we cannot provide this antibody in conjugation with Alexa Fluor or FITC in fact with any conjugate. Sorry t...

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To our knowledge, ab2792 has not been tested in drosophila. Therefore, I can offer a discount off a future purchase if you buy ab2792 now, test it in drosophila and submit feedback to us in th...

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To our knowledge, ab2792 has only been tested in Rat, Hamster, Dog, Human and Monkey and it is predicted to react with species such as Fruit fly because the homology between the Rat PDI (used as...

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I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab2792.

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Thank you for your response. This is to let you know that I have placed a new order for you - for one vial of ab2792 as a free of charge replacement and the new order number is 11140. I hope the second vial will work as it is expected, and pleas...

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Thank you for your email. We have checked with the source of the antibody through their development records for an answer to this question, but we were unsuccessful - it just says PDI rather than a specific isoform. The original reference for this ...

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Thank you for your enquiry. As we discussed, you should be using an anti-IgG2a secondary antibody. Please try this and if it does not work then please let me know. Here is the isotype: Isotype IgG2a. Here also is the datasheet: PDI antib...

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Thank you for your enquiry. I found some additional information regarding the use of this antibody in IF. This antibody has been tested briefly in Immunofluorescence in U20S cells (human). IF was performed with a standard paraformaldehyde technique ...

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